The promoter used in this product is CVM which is designed for use with Mammalian systems. This product has not been tested outside of Mammalian systems.
Pricing and availability is not currently available.
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recombinant
expressed in E. coli
Quality Level
packaging
vial of 50 μL
concentration
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
shipped in
dry ice
storage temp.
−20°C
General description
This product is an expression plasmid that utilizes the CMV promoter for strong transient expression of eSpCas9 and GFP linked by a 2A peptide (CMV-eSpCas9-2A-GFP) allowing for easy visualization of successful transfection. The eSpCas9 expression plasmid is one part of a two part CRISPR system with individual eSpCas9 and gRNA expression vectors.
To order gRNA in any format click here
To order gRNA in any format click here
Application
Functional Genomics/Target Validation
- Creation of gene knockouts in multiple cell lines
- Complete knockout of genes not amenable to RNAi
- Creation of knock-in cell lines with promoters, fusion tags, or reporters integrated into endogenous genes
Features and Benefits
- Enhanced specificity compared to wild type Cas9
- Highly Active
- Ready to use purified plasmid DNA
Principle
CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB. Newly engineered eSpCas9 enables the efficient targeted gene editing of established CRISPR systems with the benefit of reduced off-target effects. Point mutations in the chromosome-binding motif of SpCas9, as described by Slaymaker, et al., provide higher on-target fidelity without loss of cleavage efficiency.[1]
Legal Information
Storage Class Code
10 - Combustible liquids
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
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Certificates of Analysis (COA)
Lot/Batch Number
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Rationally engineered Cas9 nucleases with improved specificity.
Slaymaker, I.M., et al.
Science, 351, 84-88 (2015)
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Is this plasmid suitable for bacteria amplification and metazoan GFP expression?
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